Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells
We examined what effects are exerted by expression of the bcl-2 gene and by treatment with nerve growth factor (NGF) on the intensity of apoptosis in cultured pheochromocytoma cells (PC12 cells). Half of these cells were transduced with the bcl-2 gene using lentiviral plasmids, and the respective...
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| Дата: | 2014 |
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| Автори: | , , , , |
| Формат: | Стаття |
| Мова: | English |
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Інститут фізіології ім. О.О. Богомольця НАН України
2014
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| Назва видання: | Нейрофизиология |
| Онлайн доступ: | http://dspace.nbuv.gov.ua/handle/123456789/148307 |
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| Цитувати: | Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells / G. Han, W. Wei, X. Zhang, Zh. Lai, Ch. Chen // Нейрофизиология. — 2014. — Т. 46, № 4. — С. 375-380. — Бібліогр.: 11 назв. — англ. |
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irk-123456789-1483072019-02-18T01:26:07Z Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells Han, G. Wei, W. Zhang, X. Lai, Zh. Chen, Ch. We examined what effects are exerted by expression of the bcl-2 gene and by treatment with nerve growth factor (NGF) on the intensity of apoptosis in cultured pheochromocytoma cells (PC12 cells). Half of these cells were transduced with the bcl-2 gene using lentiviral plasmids, and the respective two groups were denoted as bcl-2-PC12 and control (c) PC12. Then the cells were incubated in a serum-free medium in six different modes. One group of c-PC12 cells was incubated in this medium with no additional agents added, another group was incubated with 1.0 mM H₂O₂ , and the third group was incubated with both 1 mM H₂O₂ and 20 ng/ml NGF (groups 1-3). Cells of the another triad were incubated under the same conditions, respectively, but these were bcl-2-PC12 cells (groups 4-6). The apoptosis rate in each group after 1-h-long incubation was measured using a flow cytometry method. A bicinchoninic acid (BCA) technique was used for estimation of expression of Bcl-2 protein in the cultures. As was observed, the action of H₂O₂ significantly increased the apoptosis rate in both c-PC12 and bcl-2-PC12 samplings, while simultaneous action of NGF considerably attenuated such increases. At the same time, values of the apoptosis rate for bcl-2-PC12 cells were much smaller than the respective values for c-PC12 cells under all the three modes of incubation. In H₂O₂ -treated cultures, the amount of Bcl-2 protein dropped, while the treatment with NGF counteracted such shifts. The content of this protein in the bcl-2-PC12 groups was much higher than in the c-PC12 groups. Thus, transduction with the bcl-2 gene significantly inhibits apoptosis in cultured PC12 cells, and a combined influence of expression of this gene and treatment with NGF produces a synergistic effect. Ми досліджували впливи експресії гена bcl-2 та дії нервового фактора росту (NGF) на інтенсивність апоптозу культивованих клітин феохромоцитоми (PC12). У половину таких клітин був трансдукований ген bcl-2; відповідні дві групи зразків були позначені як bcl-2-PC12 та контрольні (с-PC12). Потім шість груп клітин інкубували в безсироватковому середовищі в різних умовах. Перша група клітин с-PC12 інкубувалася без дії будь-яких додаткових агентів, друга група – з додаванням 1 мМ H₂O₂ , а третя – в присутності як 1 мМ H₂O₂ , так і 20 нг/мл NGF. Клітини трьох інших груп (4–6) інкубували в тих самих умовах, але це були клітини bcl-2-PC12. Ступінь апоптозу в кожній групі після одногодинної інкубації вимірювали з використанням методу флоуцитометрії. Методику з використанням біцинхонінової кислоти застосовували для оцінки експресії білка Bcl-2 в культурах. Як було виявлено, дія H₂O₂ істотно збільшувала ступінь апоптозу в зразках як c-PC12, так і bcl-2-PC12, але одночасна дія NGF помітно зменшувала таке зростання. В той же час інтенсивності апоптозу клітин bcl-2-PC12 були значно меншими, ніж відповідні значення у клітин c-PC12 при всіх трьох режимах інкубації. В культурах, підданих впливу H₂O₂ , кількість протеїну Bcl-2 була зменшеною, тоді як вплив NGF протидіяв таким зрушенням. Вміст згаданого протеїну в групах bcl-2-PC12 був значно вищим, ніж у групах c-PC12. Отже, трансдукція гена bcl2 істотно гальмує апоптоз культивованих клітин PC12, а комбінований вплив експресії цього гена та аплікації NGF забезпечує сінергічні ефекти. 2014 Article Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells / G. Han, W. Wei, X. Zhang, Zh. Lai, Ch. Chen // Нейрофизиология. — 2014. — Т. 46, № 4. — С. 375-380. — Бібліогр.: 11 назв. — англ. 0028-2561 http://dspace.nbuv.gov.ua/handle/123456789/148307 612.014.3+576.385.4 en Нейрофизиология Інститут фізіології ім. О.О. Богомольця НАН України |
| institution |
Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| collection |
DSpace DC |
| language |
English |
| description |
We examined what effects are exerted by expression of the bcl-2 gene and by treatment
with nerve growth factor (NGF) on the intensity of apoptosis in cultured pheochromocytoma
cells (PC12 cells). Half of these cells were transduced with the bcl-2 gene using lentiviral
plasmids, and the respective two groups were denoted as bcl-2-PC12 and control (c) PC12.
Then the cells were incubated in a serum-free medium in six different modes. One group of
c-PC12 cells was incubated in this medium with no additional agents added, another group
was incubated with 1.0 mM H₂O₂
, and the third group was incubated with both 1 mM H₂O₂
and 20 ng/ml NGF (groups 1-3). Cells of the another triad were incubated under the same
conditions, respectively, but these were bcl-2-PC12 cells (groups 4-6). The apoptosis rate
in each group after 1-h-long incubation was measured using a flow cytometry method. A
bicinchoninic acid (BCA) technique was used for estimation of expression of Bcl-2 protein
in the cultures. As was observed, the action of H₂O₂ significantly increased the apoptosis rate
in both c-PC12 and bcl-2-PC12 samplings, while simultaneous action of NGF considerably
attenuated such increases. At the same time, values of the apoptosis rate for bcl-2-PC12 cells
were much smaller than the respective values for c-PC12 cells under all the three modes of
incubation. In H₂O₂
-treated cultures, the amount of Bcl-2 protein dropped, while the treatment
with NGF counteracted such shifts. The content of this protein in the bcl-2-PC12 groups was
much higher than in the c-PC12 groups. Thus, transduction with the bcl-2 gene significantly
inhibits apoptosis in cultured PC12 cells, and a combined influence of expression of this gene
and treatment with NGF produces a synergistic effect. |
| format |
Article |
| author |
Han, G. Wei, W. Zhang, X. Lai, Zh. Chen, Ch. |
| spellingShingle |
Han, G. Wei, W. Zhang, X. Lai, Zh. Chen, Ch. Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells Нейрофизиология |
| author_facet |
Han, G. Wei, W. Zhang, X. Lai, Zh. Chen, Ch. |
| author_sort |
Han, G. |
| title |
Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells |
| title_short |
Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells |
| title_full |
Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells |
| title_fullStr |
Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells |
| title_full_unstemmed |
Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells |
| title_sort |
effects of transduction of the bcl-2 gene and of nerve growth factor on apoptosis of cultured pc12 cells |
| publisher |
Інститут фізіології ім. О.О. Богомольця НАН України |
| publishDate |
2014 |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/148307 |
| citation_txt |
Effects of Transduction of the bcl-2 Gene and of Nerve Growth Factor on Apoptosis of Cultured PC12 Cells / G. Han, W. Wei, X. Zhang, Zh. Lai, Ch. Chen // Нейрофизиология. — 2014. — Т. 46, № 4. — С. 375-380. — Бібліогр.: 11 назв. — англ. |
| series |
Нейрофизиология |
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2023-05-20T17:30:00Z |
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